Seasonal variations in circulating levels of progesterone and estradiol in unmated adult female woodchucks (Marmota monax) in captivity.

The annual profile of serum levels of progesterone (P) and estradiol-17 beta (E2) was characterized in a seasonally breeding rodent, the woodchuck (Marmota monax). Hormonal levels were determined in serum samples taken at weekly or biweekly intervals from unmated female woodchucks maintained all year indoors under controlled conditions of photoperiod and temperature. Annual fluctuations included a rise of E2 levels during late January through February, followed by a modest increase in plasma P concentrations by late March, the latter attaining peak values during April and May. A temporal dissociation of peak values of circulating levels of P and E2 during the annual reproductive cycle was also detected. The timing of changes in serum levels of P and E2 in these captive woodchucks corresponded to reproductive events during the normal breeding season of the woodchuck in the southern part of its range.     

Metabolism of phenylalanine in mice homozygous for the gene `dilute lethal''

Mice homozygous for d(l) have been suggested as models for phenylketonuria. We found: (1) the concentration of phenylalanine in the blood was normal at all ages examined; (2) phenylalanine hydroxylase activity in the liver in vitro equalled that in unaffected littermates; (3) the apparent K(m) values for phenylalanine and cofactor respectively in d(l)/d(l) mice were the same as in their normal littermates; (4) inhibition of the overall reaction by the particulate fraction, excess of substrate, excess of cofactor or phenylpyruvic acid showed no difference between d(l)/d(l) mice and their unaffected littermates; (5) phenylalanine injected in vivo had equal, small, effects on phenylalanine hydroxylase activity of the liver measured in vitro in the two groups of mice. An explanation of the findings of other workers, based on the natural history of the disease process, is tentatively put forward.     

Predicting properties of intrinsically unstructured proteins

There is increasing evidence that intrinsically unstructured proteins or protein domains have important biological functions. These types of proteins may be productively analyzed using polymer theory developed to predict global physical properties of polymers. In these theories molecular detail is “coarse grained” out of the models, and replaced with a small number of parameters that characterize the polymer. This reduction in complexity allows extremely large systems to be studied. In the case of simulations, the time scales accessible also increase significantly. Here we discuss the application of polymer theory to unstructured proteins, and consider how to classify proteins within a polymer framework. We then review polymer theory that is relevant to predicting functionally important properties, such as radius of gyration, height of a polymer brush and force required to compress a polymer brush.     

Determination of rofecoxib, a cyclooxygenase-2 specific inhibitor, in human plasma using high-performance liquid chromatography with post-column photochemical derivatization and fluorescence detection

A method for the determination of rofecoxib in human plasma is described. After the addition of an internal standard, buffered (pH 5) plasma samples are extracted with hexane–methylene chloride (50:50, v/v). The extracts are evaporated to dryness and reconstituted in mobile phase. Upon exposure to UV light, the analyte was found to undergo a stilbene–phenanthrene-like photocyclization reaction with the resulting formation of a highly fluorescent species. Thus, the plasma extracts were analyzed via HPLC with post-column photochemical derivatization and fluorescence detection. The assay has been validated in the concentration range of 0.5–100 ng/ml using 1-ml samples. The method has been successfully utilized to support human clinical pharmacokinetic studies.